JP2023550594A - GLP-1, GIP, and glucagon receptor triple agonist - Google Patents

Abstract

The present invention relates to peptides and derivatives thereof that are GLP-1/GIP/glucagon receptor triple agonists and describes their medical use in the treatment and/or prevention of obesity, diabetes, and/or liver diseases.

Description

The present invention provides novel compounds that are agonists of the glucagon-like peptide 1 receptor (GLP-1R), the glucose-dependent insulinotropic polypeptide receptor (GIPR), and the glucagon receptor (GcgR) with a prolonged action profile. Regarding.

INCORPORATION BY REFERENCE OF THE SEQUENCE LISTING This application is filed with a Sequence Listing in electronic form. The entire contents of the Sequence Listing are incorporated herein by reference. The sequence listing entitled “200078WO01 sequence listing_ST25” is 37,918 bytes and was created on October 21, 2021.

Glucagon-like peptide 1 (GLP-1) is a hormone derived from enteroendocrine cells and is one of the two prominent endogenous physiological incretins. GLP-1 improves glycemic control by stimulating glucose-dependent insulin secretion in response to nutrients (glucose), inhibits glucagon secretion from pancreatic alpha cells, slows gastric emptying, and primarily Induces weight loss by reducing food consumption. Another prominent incretin, glucose-dependent insulinotropic polypeptide (GIP), improves glycemic control by stimulating insulin secretion in response to nutrients (fat, glucose). Furthermore, GIP is thought to improve plasma lipid profile and stimulate calcium accumulation in bone. GIP analogs induce weight loss and improve glycemic control in dual administration and by additive/synergistic effects with GLP-1 analogs as single molecule co-agents (Non-Patent Document 1, Non-patent Document 2, Non-patent Document 3, Non-patent Document 4), it is a suitable candidate for amplification of GLP-1-based pharmacology.

Glucagon (Gcg) increases hepatic glucose production by promoting glycogenolysis and gluconeogenesis and prevents the occurrence of hypoglycemia. In addition, glucagon has been shown to stimulate lipolysis, increase energy expenditure, and decrease hepatic lipoprotein production. Glucagon has been shown to provide further weight loss when combined with GLP-1 than that achieved with GLP-1 alone (Non-Patent Reference 5, Non-Patent Reference 6, Non-Patent Reference 7). . This is thought to be driven by the anorexigenic/satiety effects of GLP-1 combined with the thermogenic and lipolytic effects of glucagon. The insulinotropic and insulin-sensitizing effects of GLP-1 offset the potential diabetogenic effects of chronic glucagon action, as shown in clinical studies of two GLP-1/Gcg receptor co-agonists. (Non-patent Document 8, Non-Patent Document 9). However, the ratio of GLP-1 activity to glucagon activity needs to be carefully managed to maintain glycemic control, which limits the achievable weight-lowering effects.

Glucagon's limited therapeutic window may be alleviated by combining both GLP-1 and GIP, both of which acutely stimulate glucose-dependent insulin secretion and chronically enhance insulin sensitivity. This dual incretin activity is predicted based on murine pharmacology and allows for a more aggressive glucagon pharmacology, driving additional efficacy with reduced diabetogenic propensity. In addition, glucagon's effects on energy expenditure are not offset by GLP-1 and GIP, which provide their own appetite suppressant effects. Therefore, triple agonist peptides are expected to provide glucose management with greater fat mass loss than can be achieved with either GLP-1/GIP or GLP-1/Gcg co-agonism. (Non-Patent Document 10).

GLP-1/GIP/Gcg receptor triple agonists and their potential medical uses have been described in several patent applications. Prolonged triple agonists suitable for weekly administration in humans are described in US Pat. Co-administration of prolonged glucagon analogs and prolonged GLP-1/GIP co-agonists is described in US Pat. Triple agonists designed for prolongation via conjugation to Fc fragments are described in US Pat. Exendin-4 derivatives with triple agonistic activity and prolongation suitable for once-daily administration in humans are described in US Pat. There is. However, neither triple agonist product has ever received marketing approval.

International Publication No. 2019/125929 International Publication No. 2019/125938 International Publication No. 2015/067716 International Publication No. 2017/009236 US Patent Publication No. 2019/0218269 US Patent Publication No. 2019/0153060 International Publication No. 2015/155141 International Publication No. 2014/096145 International Publication No. 2014/096148 International Publication No. 2014/096150

Finan et al, Sci Transl Med, 2013, 5(209): 209ra151 Frias et al, Cell Metab, 2017, 26(2): 343-352 Coskun et al, Mol Metab, 2018, 18:3-14 Frias et al, Lancet, 2018, 392(10160): 2180-2193 Evers et al, J Med Chem, 2017, 60(10): 4293-4303 Henderson et al, Diabetes Obes Metab 2016, 18(12): 1176-1190 Day et al, Nat Chem Biol, 2009, 5(10): 749-757 Tillner et al, Diabetes Obes Metab, 2019, 21(1): 120-128 Parker et al, J Clin Endocrinol Metab, 2019, 105(3): 803-820 Finan et al, Nat. Med. , 2015, 21(1): 27-36

The present invention relates to a GLP-1/GIP/Gcg receptor triple agonist comprising a peptide and a substituent, wherein the substituent is covalently bonded to the peptide at a position selected from the group consisting of positions 16, 17, and 21. For example, the substituent is linked via the epsilon-amino group of a lysine at a position selected from the group consisting of positions 16, 17, and 21 of the peptide. This substituent enhances the half-life of the triple agonist and includes a difatty acid containing protractor A- and a linker BC-. The invention also relates to pharmaceutical compositions comprising such triple agonists and pharmaceutically acceptable excipients, and medical uses of the triple agonists.

In one aspect, the invention provides a GLP-1/GIP/Gcg receptor triple agonist comprising a peptide and a substituent, wherein the amino acid sequence of the peptide is
X ₁ X ₂ X _{3 GTFTSDYS 12 X 13 LX 15} KX _{17 X 18} AX 20
X ₁ is H or Y,
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is K, I, or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
X ₂₈ is A or E,
the peptide is a C-terminal acid or amide;
a GLP-1/GIP/Gcg receptor triple agonist, wherein the substituent is covalently bonded to the epsilon-amino group of K at a position selected from the group consisting of positions 16, 17, and 21;
or a pharmaceutically acceptable salt thereof.

Additionally or alternatively, in a second aspect, the invention provides a GLP-1/GIP/Gcg receptor triple agonist and, optionally, at least one pharmaceutically acceptable excipient. A pharmaceutical composition comprising:

Additionally or alternatively, in a further aspect, the present invention relates to a GLP-1/GIP/Gcg receptor triple agonist as described herein for use as a medicament.

Additionally or alternatively, in a further aspect, the present invention provides a GLP-1/GIP/Gcg receptor as described herein for use in the prevention and/or treatment of diabetes, obesity, and/or liver disease. Concerning triple agonists.

Figure 1 shows the time course of the maximum efficacy study in DIO mice comparing vehicle (-□-), Compound No. 21 (-●-), Compound No. 30 (-△-), and Compound No. 33 (-▼-). It shows the change in weight. DIO mice received a once daily subcutaneous dose according to the titration schedule listed in Table 4. Figure 2 shows the cumulative food intake in a maximum efficacy study in DIO mice comparing vehicle (-□-), Compound No. 21 (-●-), Compound No. 30 (-△-), and Compound No. 33 (-▼-). Indicates intake. DIO mice received a once daily subcutaneous dose according to the titration schedule listed in Table 4.

The present invention relates to compounds that are agonists of the respective receptors GLP-1, GIP, and glucagon.

In the following, Greek letters may be represented by their symbols or corresponding names, for example α is alpha, β is beta, ε is epsilon, γ is gamma, ω is omega. It is as follows. Also, the Greek letter for μ may be represented by "u", for example, μl is ul and μM is uM.

Unless expressly stated otherwise herein, terms appearing in the singular include the plural.

Also described herein are triple agonists, pharmaceutical compositions, and uses thereof, and non-limiting terms such as "comprising" and "comprising" are used to describe Replaced with an exclusive term such as "

GLP-1/GIP/Glucagon Receptor Triple Agonists The present invention relates to compounds that are agonists of GLP-1 receptors, GIP receptors, and glucagon receptors. Each of these agonists can also be referred to as a "GLP-1/GIP/Gcg receptor triple agonist" or a "triple agonist." Receptor agonists, as described herein, are compounds that bind to a receptor and elicit a natural ligand response (e.g., “Principles of Biochemistry”, AL Lehninger, DL Nelson, MM Cox, Second Edition, Worth (See Publishers, 1993, page 763.)

In some embodiments, a GLP-1/GIP/Gcg triple agonist is a compound that binds to each of the three receptors GLP-1R, GIPR, and GcgR and elicits a response at each receptor; It is a compound that can activate each of the body's GLP-1R, GIPR, and GcgR.

The term "compound" is used herein to refer to a molecular entity, and thus a "compound" may have different structural elements other than the minimum elements defined for each compound or group of compounds. A compound may be a peptide or a derivative thereof as long as it contains defined structural and/or functional elements.

The term "compound" is also meant to include its pharmaceutically relevant forms, ie, a compound as defined herein or a pharmaceutically acceptable salt, amide, or ester thereof.

The term "peptide" refers to a sequence of two or more amino acids. A "peptide" may include amino acid extensions at the N-terminal and/or C-terminal positions, and/or truncations at the N-terminal and/or C-terminal positions. Generally, amino acid residues can be identified by their full name, their one letter code, and/or their three letter code. These three methods are completely equivalent.

Amino acids are molecules that contain an amino group and a carboxylic acid group, and optionally one or more additional groups, often called side chains.

The term "amino acid" includes not only proteinogenic (or natural) amino acids (particularly the 20 standard amino acids), but also non-proteinogenic (or non-natural) amino acids. Proteinogenic amino acids are those that are naturally incorporated into proteins. Standard amino acids are amino acids encoded by the genetic code. Non-proteinogenic amino acids are either not found in proteins or are not produced by standard cellular machinery (eg, they may not be subjected to post-translational modification). Non-limiting examples of non-proteinogenic amino acids are Aib (α-aminoisobutyric acid, or 2-aminoisobutyric acid), norleucine (Nle), norvaline, and the D isomer of proteinogenic amino acids.

In the following, each amino acid of a peptide for which the optical isomer is not stated is to be understood as meaning the L isomer (unless specified otherwise).

The GLP-1/GIP/Gcg triple agonists described herein include or consist of a peptide and a substituent. In some embodiments, the peptide is a synthetic peptide engineered to optimize activity at GLP-1, GIP, and Gcg receptors. Compounds with receptor activity for each of the GLP-1, GIP, and Gcg receptors have been identified as shown in the Examples herein.

The triple agonists described herein further exhibit extended half-lives afforded by fatty acid-containing substituents.

In some embodiments, the carboxy terminus of the triple agonist carries a carboxylic acid group (-COOH), also referred to as the C-terminal acid. In some embodiments, the carboxy terminus of the triple agonist may optionally include an amide group (C(=O)-NH ₂ ), also referred to as a C-terminal amide, as found in native exendin-4. This is a naturally occurring modification in some proteins that replaces -OH with _-NH2 .

Peptides The GLP-1/GIP/Gcg receptor triple agonists described herein include peptides and substituents, where the substituents are arranged via amino acid residues, e.g., via functional groups on amino acid side chains. and is bonded to the peptide backbone. In some embodiments, a triple agonist described herein consists of a peptide and a substituent.

In some embodiments, _the amino acid sequence of _the peptide is X ₁ X ₂ X _{3 GTFTSDYSX 12} X ₁₃ LX ₁₅ KX ₁₇ X ₁₈ AX ₂₀
X ₁ is H or Y,
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is K, I, or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
_X28 is A or E,
The peptide is a C-terminal acid or amide.

In some _embodiments , _the amino acid sequence _{of the peptide is HX2HGTFTSDYSX12X13LX15KX17X18AX20EFVX24WLLX28GGPSSGAPPPS} (SEQ _ID NO: ₄₇ );
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
X ₂₈ is A or E.

In some embodiments, the amino acid sequence of the peptide is YX ₂ QGTFTSDYSX ₁₂ X ₁₃ LX ₁₅ KX ₁₇ X ₁₈ AX ₂₀ EFVX ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO:50);
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
X ₂₈ is A or E.

In some embodiments, X ₁ X ₂ X ₃ is selected from the group consisting of HAibH and YAibQ. In some embodiments, X ₁ X ₂ X ₃ is HAibH. In some embodiments, X ₁ X ₂ X ₃ is YAibQ.

In some embodiments, X ₁₂ is I or Y. In some embodiments, X ₁₅ is D. In some embodiments, X ₁₅ is E. _In some embodiments, _{SX 12} selected from the group consisting of. In some embodiments, SX ₁₂ X ₁₃ LX ₁₅ is selected from the group consisting of SYYLE, SILLE, SIYLE, and SIYLD. In some embodiments, SX ₁₂ X ₁₃ LX ₁₅ is selected from the group consisting of SYYLE and SILLE.

In some _{embodiments ,} KX ₁₇ . In some embodiments, KX ₁₇ X ₁₈ AX ₂₀ is KQAAAib.

In some embodiments, _X24 is N. In some embodiments, _X24 is Q.

In some embodiments, _X28 is A. In some embodiments, _X28 is E.

In some embodiments, the peptide is selected from any one of SEQ ID NOs: 1-33. In some embodiments, the amino acid sequence of the peptide is selected from the group consisting of SEQ ID NOs: 1, 11, 13-14, 17-18, 20, and 26. In some embodiments, the amino acid sequence of the peptide is selected from the group consisting of SEQ ID NOs: 11, 14, and 26.

In some embodiments, the peptide is a C-terminal acid or amide. In some embodiments, the peptide is a C-terminal acid. In some embodiments, the peptide is a C-terminal amide.

Derivatives In some embodiments, the GLP-1/GIP/Gcg receptor triple agonist comprises a substituent covalently attached to the peptide. Such compounds are sometimes referred to as derivatives of peptides, as they are obtained by covalently bonding substituents to the peptide backbone, for example via functional groups on the amino acid side chains. In some embodiments, the GLP-1/GIP/Gcg receptor triple agonist consists of a peptide and a substituent.

One aspect of the present _invention is a compound comprising _{a peptide} and _{a substituent ,} wherein _the amino acid _{sequence of the} peptide _{is :} SEQ ID NO: 45),
X ₁ is H or Y,
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is K, I, or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
X ₂₈ is A or E,
the peptide is a C-terminal acid or amide;
It relates to a compound, or a pharmaceutically acceptable salt thereof, wherein the substituent is covalently bonded to the epsilon-amino group of K at a position selected from the group consisting of the 16th, 17th, and 21st positions.

In further embodiments, the peptide may be defined as herein above.

Substituents In some embodiments, a substituent is covalently attached to the peptide backbone of a triple agonist described herein, for example, via the epsilon-amino group of a lysine (K) residue. In some embodiments, the substituent is covalently attached through the epsilon-amino group of lysine (K) at a position selected from the group consisting of positions 16, 17, and 21 of the backbone. In a preferred embodiment, the substituent is covalently bonded through the epsilon-amino group of lysine (K) at position 16.

In one aspect, the substituent is capable of forming a non-covalent complex with a protein, e.g., albumin or serum albumin, thereby facilitating circulation of the triple agonist in the bloodstream, and also The albumin complex also has the effect of prolonging the action time of the triple agonist, since it is not slowly removed by renal clearance.

In some embodiments, the substituent is attached to a lysine residue of the peptide backbone by acylation, i.e., through an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine. covalently bonded to In some embodiments, the amino group of the lysine is linked to the substituent aldehyde by reductive amination.

In some embodiments, the substituent is ABC- and includes a protractor (A-) and a linker element (B-C-). In some embodiments, the protractor is a fatty acid and is the terminal moiety of a substituent responsible for increasing the half-life of the compound.

In some embodiments, the protractor, A-, has the following formula 1:
Chemical formula 1: HOOC-(CH ₂ ) _p -CO-*
where p is an integer ranging from 8 to 20. In some embodiments, A is Formula 1 and p is an integer ranging from 14 to 20, such as 16 or 18. Formula 1 is also sometimes referred to as C(n+2) diacid.

In some embodiments, the substituent further comprises a linker element BC-, where B- is selected from Formula 2 or Formula 3 below,
Chemical formula 2: *-NH-CH(COOH)-(CH ₂ ) ₂ -CO-*
Formula 2 can also be called γGlu,
and includes both the R- and S-forms. In some embodiments, Formula 2 is the S-isomer of γGlu.
Formula 3: *-NH-CH ₂ -(C ₆ H ₁₀ )-CO-NH-CH(COOH)-(CH ₂ ) ₂ -CO-* can also be described by Formula 3a or 3b below,
Formula 3 may also be referred to as Trx-γGlu, where Trx refers to tranexamic acid or trans-4-(aminomethyl)cyclohexanecarboxylic acid, and formula 3 is (1,2), (1,3), and (1,4) forms, while Formula 3a and Formula 3b specify the (1,4) form.

In some embodiments, the substituent further comprises a linker element that can be defined by BC-, where C- is selected from Formula 4 or Formula 5 below,
Chemical formula 4: *-[NH-((CH ₂ ) ₂ -O) ₂ -CH ₂ -CO] ₂ -*, which is two 8-amino-3,6-dioxas linked via an amide bond. It may also be called Ado-Ado, which consists of an octanoic acid moiety.
Formula 5: *-[NH-( _CH2 ) ₄ -CH( _NH2 )-CO] _2- *, which also consists of two epsilonlysine moieties linked via an amide bond. It may also be called εLys-εLys,
and includes both the R- and S-forms. In one embodiment, Formula 5 is the S-isomer of both εLys moieties.

The symbol * indicates the point of attachment, and A-, B-, and C- are interconnected by amide bonds in the order shown.

In some embodiments, the substituent ABC- is
and
selected from the group consisting of.

In some embodiments, the substituents are selected from the group consisting of Formula 6, Formula 7, Formula 12, and Formula 13. In certain embodiments, substituent ABC- is selected from Formula 12 or Formula 13.

Pharmaceutically Acceptable Salts In some embodiments, the triple agonists described herein are in the form of pharmaceutically acceptable salts.

Salts are formed, for example, by a chemical reaction between a base and an acid, such as 2NH ₃ +H ₂ SO ₄ →(NH ₄ ) ₂ SO ₄ .

Salts may be basic salts, acidic salts, or neither (ie, neutral salts). In water, basic salts produce hydroxide ions and acidic salts produce hydronium ions.

In some embodiments, the triple agonist salts of the invention are formed by the addition of a cation or anion between anionic or cationic groups, respectively. These groups may be located on the peptide moiety and/or the side chain of the triple agonist.

Non-limiting examples of anionic groups of triple agonists described herein include free carboxyl groups on side chains and free carboxyl groups on peptide moieties. In some embodiments, the peptide moiety includes a free carboxylic acid group at the C-terminus. In some embodiments, the peptide moiety includes free carboxylic acid groups on internal amino acid residues such as Asp and Glu. In some embodiments, the triple agonists described herein do not contain free carboxylic acid groups.

Non-limiting examples of cationic groups on peptide moieties include free amino groups at the N-terminus and on internal basic amino acid residues such as His, Arg, and Lys. In some embodiments, the triple agonists described herein do not include a free amino group at the N-terminus.

In certain embodiments, the triple agonist is in the form of a pharmaceutically acceptable salt.

Functional Properties In a first aspect, the triple agonists described herein include all of the GLP-1R, GIPR, and GcgR agonists tested by in vitro efficacy as described in Example 2 herein. It is an agonist in Thus, the triple agonists described herein are capable of activating each of GLP-1R, GIPR, and GcgR.

Additionally or alternatively, in a second functional aspect, the triple agonist has in vivo efficacy on body weight, food intake, and glucose tolerance.

Additionally or alternatively, in a third functional aspect, the triple agonist produces improved pharmacokinetic properties over the natural hormones GLP-1, GIP, and Gcg.

Biological Activity - In Vitro Efficacy In this section, the term "potency" is used interchangeably with "biological activity" and "activity."

According to a first functional aspect, the triple agonists described herein are potent at GLP-1R, GIPR, and GcgR. The peptides described herein are also potent at GLP-1R, GIPR, and GcgR. In some embodiments, the triple agonists described herein have the potency of in vitro efficacy to activate hGLP-1R, hGIPR, and hGcgR.

In some embodiments, efficacy is determined by the in vitro efficacy of a triple agonist described herein (or herein) in a functional receptor assay for each of GLP-1R, GIPR, and GcgR. determined by the performance of the described peptide). In some embodiments, the functional receptor assay determines the in vitro efficacy of a triple agonist described herein (or a peptide described herein) for each of hGLP-1R, hGIPR, and hGcgR. be determined individually.

The term "half-maximal effective concentration" (also known as "EC ₅₀ ") refers to the concentration that induces a response midway between baseline and maximum, by reference to a dose-response curve. The EC ₅₀ is used as a measure of a compound's potency and represents the concentration at which 50% of its maximal effect is observed.

Accordingly, the in vitro efficacy of the triple agonists described herein can be determined by determining the _EC50 , as described below. The lower the _EC50 , the better the efficacy. To further characterize these compounds, in vitro comparisons were made with the natural hormones of each receptor, namely hGLP-1(7-37) (SEQ ID NO: 35), hGIP (SEQ ID NO: 36), and hGcg (SEQ ID NO: 37). It may be appropriate to consider the efficacy in

Efficacy in vitro can be determined, for example, in culture containing membranes expressing the appropriate receptor and/or in assays using whole cells expressing the appropriate receptor.

For example, the functional response of a human GLP-1, GIP, or Gcg receptor can be determined in a receptor gene assay by expressing, e.g. can be measured in a stably transfected BHK cell line containing response element (CRE) DNA and the firefly luciferase (CRE luciferase) gene. The generation of cAMP as a result of activation of GLP-1, GIP, or Gcg receptors then results in the expression of luciferase. Luciferase can be determined by adding luciferin, which is converted by the enzyme to oxyluciferin to produce bioluminescence, which is measured as a reporter of potency in vitro. One non-limiting example of such an assay is described in Example 2 herein.

In some embodiments, triple agonists described herein can activate hGLP-1R, hGIPR, and hGcgR. In some embodiments, the triple agonists described herein have an EC ₅₀ of less than 500 pM, such as less than 100 pM, such as less than 50 pM, in vitro in the CRE luciferase reporter gene assay described in Example 2. It can activate hGLP-1R, hGIPR, and hGcgR.

In some embodiments, the triple agonist described herein corresponds to an EC ₅₀ of 200 pM or less, more preferably less than 75 pM, or most preferably less than 50 pM, as determined using the method of Example 2. It has in vitro efficacy at hGLP-1R, hGIPR, and hGcgR.

In one embodiment, the EC ₅₀ in the human GLP-1, GIP, and Gcg receptor assay described in Example 2 is from 0.5 to 500 pM, such as from 0.5 to 100 pM, or such as from 0.5 to 50 pM. It is.

Biological Activity in DIO Mice - Pharmacodynamic Studies According to the second functional aspect, the triple agonists described herein are biologically active in vivo, which can be achieved in any suitable animal. It can be determined by any technique known in the art in models and clinical trials.

Diet-induced obesity (DIO) mice are one example of a suitable animal model, and effects on body weight, food intake, and glucose tolerance can be evaluated during subchronic dosing in this model. The effects of the triple agonists described herein on body weight, food intake, and glucose tolerance can be determined in vivo in such mice, e.g., as described in Example 4 herein. .

In some embodiments, the triple agonists described herein exhibit the ability to reduce body weight, food intake, and improve glucose tolerance in DIO mice, as described in Example 4.

In some embodiments, a triple agonist described herein reduces body weight in DIO mice.

In some embodiments, a triple agonist described herein reduces food intake in DIO mice.

In some embodiments, the triple agonists described herein improve glucose tolerance in DIO mice.

In some embodiments, the triple agonist described herein provides at least 10%, or 3 nmol/kg of the compound after once daily administration of 1 nmol/kg of the compound for 13 days in DIO mice. Reduce body weight by at least 25% after once daily administration. In some embodiments, the triple agonist described herein provides at least 20%, or 3 nmol of the compound compared to vehicle after administering 1 nmol/kg of the compound once daily for 13 days in DIO mice. /kg of the compound once daily reduces food intake by at least 50% compared to vehicle. In some embodiments, the triple agonist was administered once daily at 1 nmol/kg or 3 nmol/kg of the compound for 13 days in DIO mice as measured by IPGTT (intraperitoneal glucose tolerance test). Improves glucose tolerance by at least 30% later compared to vehicle.

Pharmacokinetic Profile in Minipigs - In Vivo Half-Life According to the third functional aspect, the triple agonists described herein have a terminal half-life, compared to, for example, the natural hormones GLP-1, GIP, and Gcg. It has improved pharmacokinetic properties such as increased An increase in terminal half-life means that the compound in question is eliminated from the body more slowly. The triple agonists described herein are associated with an extended duration of pharmacological effect.

The pharmacokinetic properties of the triple agonists described herein can be suitably determined in vivo in pharmacokinetic (PK) studies. Such studies assess how pharmaceutical compounds are absorbed, distributed, and excreted in the body and how these processes affect the concentration of triple agonists in the body over time. It is done for.

During the discovery and preclinical stages of drug development, PK studies can be performed using animal models such as mice, rats, monkeys, dogs, or pigs. Any such model can be used to test the PK properties of the triple agonists described herein.

In such studies, animals typically receive a single dose of drug in the appropriate formulation, either intravenously (i.v.), subcutaneously (s.c.) or orally (p.o.). administered in Blood samples are taken at predetermined time points after administration, and the samples are analyzed for drug concentration in a suitable quantitative assay. Based on these measurements, the time-plasma concentration profile of the study compound is plotted and a so-called non-compartmental pharmacokinetic analysis of the data is performed.

Terminal half-life is an important parameter because a long half-life indicates that less frequent administration of the compound may be possible. In some embodiments, the terminal half-life is i.p., eg, as described in Example 3 herein. v. In vivo half-life (t ¹ / ₂ ) in minipigs after administration.

In some embodiments, the terminal half-life in minipigs of the triple agonists described herein is i.p. v. When measured after administration, it is at least 30 hours, preferably at least 40 hours, even more preferably at least 60 hours.

Production Process The triple agonists or fragments thereof described herein may be produced by classical peptide synthesis, e.g., solid phase peptide synthesis using t-Boc or Fmoc chemistry or other well-established techniques. , for example, Greene and Wuts, “Protective Groups in Organic Synthesis”, John Wiley & Sons, 1999, Florencio Zaragoza Dorwald, “Organic "Synthesis on solid Phase", Wiley-VCH Verlag GmbH, 2000, and "Fmoc Solid Phase Peptide Synthesis", Edited by W. C. Chan and P. D. See White, Oxford University Press, 2000. In some embodiments, methods for preparing triple agonists are described herein. In some embodiments, the methods for preparing triple agonists described herein include a step of solid phase peptide synthesis.

Additionally or alternatively, the triple agonists described herein can be prepared by recombinant methods, i.e., by producing a host cell containing a DNA sequence encoding the triple agonist peptide sequence and capable of expressing the peptide. may be produced by culturing in a suitable nutrient medium under conditions that permit the expression of . Non-limiting examples of suitable host cells for expression of peptides are Escherichia coli, Saccharomyces cerevisiae, and mammalian BHK or CHO cell lines.

Triple agonists described herein that include unnatural amino acids can be produced, for example, as described in the "General Preparation Methods" section of the experimental part. Or, for example, Hodgson et al: “The synthesis of peptides and proteins containing non-natural amino acids”, Chemical Society Reviews, vol. ．． 33, no. 7 (2004), p. See 422-430.

Triple agonists described herein containing substituents can be produced, for example, as described in the "General Preparation Methods" section of the experimental part. In some embodiments, substituents are constructed as part of solid phase peptide synthesis or separately constructed and attached via lysine residues after solid phase peptide synthesis.

Specific examples of methods of preparing multiple triple agonists described herein are provided below.

Pharmaceutical Compositions A pharmaceutical composition comprising a triple agonist, or a pharmaceutically acceptable salt thereof, as described herein, and optionally one or more pharmaceutically acceptable excipients. It may be prepared as known in the art.

The term "excipient" broadly refers to any ingredient other than the active therapeutic ingredient. Excipients may be inert, inactive, and/or non-medically active substances.

Excipients can serve a variety of purposes, for example as carriers, vehicles, diluents, tablet adjuvants, and/or to improve the administration and/or absorption of the active substance.

Formulations of pharmaceutically active ingredients with various excipients are known in the art and are described, for example, in Remington: The Science and Practice of Pharmacy, 19th edition (1995) and any later edition. (edition).

In some embodiments, a pharmaceutical composition comprising a triple agonist described herein is a liquid formulation, such as an aqueous formulation.

For example, liquid formulations suitable for injection can be prepared using conventional techniques of the pharmaceutical industry, optionally involving dissolving and mixing the ingredients to yield the desired end product.

In some embodiments, the triple agonist described herein is dissolved in a suitable buffer at a suitable pH so that precipitation is minimized or avoided. Injectable compositions can be sterilized, for example, by sterile filtration.

Pharmaceutical Indications A further aspect of the invention relates to a GLP-1/GIP/Gcg receptor triple agonist as described herein for use as a medicament.

In certain aspects of the invention, the GLP-1/GIP/Gcg receptor triple agonists described herein may be used for the medical treatment of:
(i) Prevention and/or treatment of eating disorders such as obesity, e.g. by reducing food intake, increasing energy expenditure, reducing weight, suppressing appetite, inducing feelings of satiety; binge eating disorder, bulimia nervosa; treatment or prevention of obesity induced by the administration of antipsychotics or steroids, delayed gastric emptying, increased physical mobility, and/or comorbidities of obesity such as osteoarthritis and/or urinary incontinence. prevention and/or treatment of diseases;
(ii) weight maintenance after successful weight loss (either drug-induced or diet and exercise), i.e. prevention of weight gain after successful weight loss;
(iii) prevention and/or treatment of all forms of diabetes, including hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (young onset adult diabetes), gestational diabetes; and/or reduction of HbA1C,
(iv) slowing or preventing the progression of diabetic diseases, such as progression of type 2 diabetes, slowing the progression of impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, slowing or preventing insulin resistance, and/or or delaying the progression from type 2 diabetes that does not require insulin to type 2 diabetes that requires insulin;
(v) prevention of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver and/or Treatment.

In some embodiments, the triple agonists described herein are for use in the prevention and/or treatment of obesity. In some embodiments, the triple agonists described herein are for use in the prevention and/or treatment of type 2 diabetes. In some embodiments, the triple agonists described herein are for use in the prevention and/or treatment of non-alcoholic steatohepatitis (NASH).

In some embodiments, the triple agonists described herein are associated with methods of preventing and/or treating obesity. In some embodiments, the triple agonists described herein are associated with methods of preventing and/or treating type 2 diabetes. In some embodiments, the triple agonists described herein are associated with methods of preventing and/or treating non-alcoholic steatohepatitis (NASH).

In some embodiments, the triple agonists described herein are associated with weight management methods. In some embodiments, the triple agonists described herein are associated with methods of anorexia. In some embodiments, the invention relates to a method for reducing food intake. In some embodiments, the triple agonists described herein are associated with methods of preventing or treating overweight in a subject.

Particular Embodiments The invention is further illustrated by the following non-limiting embodiments:
1. A GLP- ₁ /GIP/Gcg receptor _triple agonist comprising a _peptide and a substituent _{, wherein the} amino acid _{sequence of the} peptide _{is :} WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 45),
X ₁ is H or Y,
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is K, I, or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
X ₂₈ is A or E,
the peptide is a C-terminal acid or amide;
a GLP-1/GIP/Gcg receptor triple agonist, wherein the substituent is covalently bonded to the epsilon-amino group of K at a position selected from the group consisting of positions 16, 17, and 21;
or a pharmaceutically acceptable salt thereof.
2. _{The amino acid} sequence _{of the peptide} is X _{1 X 2} X _{3 GTFTSDYS 12}
X ₁ is H or Y,
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
X ₂₈ is A or E,
the peptide is a C-terminal acid or amide;
GLP-1/GIP/ according to embodiment 1, wherein the substituent is covalently bonded to the epsilon amino group of K at one of the positions selected from the group consisting of positions 16, 17, and 21. Gcg receptor triple agonist,
or a pharmaceutically acceptable salt thereof.
3. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁ X ₂ X ₃ is selected from the group consisting of HAibH and YAibQ.
4. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁ X ₂ X ₃ is HAibH.
5. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁ X ₂ X ₃ is YAibQ.
6. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁₂ is I.
7. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-5, wherein X ₁₂ is Y.
8. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁₃ is Y.
9. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-7, wherein X ₁₃ is L.
10. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁₅ is D.
11. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-9, wherein X ₁₅ is E.
12. GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein SX ₁₂ X ₁₃ LX ₁₅ is selected from the group consisting of SYYLE, SILLE, SYLLE, SIYLE, and SIYLD .
13. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein SX ₁₂ X ₁₃ LX ₁₅ is selected from the group consisting of SYYLE, SILLE, SIYLE, and SIYLD.
14. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-9 or 11-13, wherein SX ₁₂ X ₁₃ LX ₁₅ is selected from the group consisting of SYYLE and SILLE.
15. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁₇ is K.
16. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-14, wherein X ₁₇ is Q.
17. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-14, wherein X ₁₇ is R.
18. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₁₈ is A.
19. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-17, wherein X ₁₈ is Y.
20. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-17, wherein X ₁₈ is K.
21. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₂₀ is Aib.
22. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-20, wherein X ₂₀ is Q.
23. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-20, wherein X ₂₀ is E.
24. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-20, wherein X ₂₀ is R.
25. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-20, wherein X ₂₀ is H.
26. GLP-1/GIP/ according to any one of embodiments 1-16, 18-19, or 21, wherein KX ₁₇ X ₁₈ AX ₂₀ is selected from the group consisting of KQAAAib, KKAAAib, KQYAAib, and KKYAAib. Gcg receptor triple agonist.
27. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₂₁ is E.
28. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-26, wherein X ₂₁ is K.
29. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₂₄ is Q.
30. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-28, wherein X ₂₄ is N.
31. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein X ₂₈ is E.
32. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-30, wherein X ₂₈ is A.
33. The amino acid sequence of the peptide is HX ₂ HGTFTSDYS ₁₂ X ₁₃ LX ₁₅ KX ₁₇ X ₁₈ AX ₂₀ EFVX ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 47),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
GLP-1/GIP/Gcg receptor according to any one of embodiments 1-4, 6-16, 18-19, 21, 26-27, or 29-32, wherein X ₂₈ is A or E. Triple agonist.
34. The amino acid sequence of the peptide is HX ₂ HGTFTSDYSX ₁₂ X ₁₃ LEKX ₁₇ X ₁₈ AX ₂₀ EFVX ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 48),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
GLP-1/ according to any one of embodiments 1-4, 6-9, 11-16, 18-19, 21, 26-27, or 29-33, wherein X ₂₈ is A or E. GIP/Gcg receptor triple agonist.
35. _The amino acid _{sequence of the} peptide is _{HX2HGTFTSDYSX12X13LEKX17X18AX20EFVQWLLEGGPSSGAPPPS} (SEQ ID NO _: 49),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
GLP- according to any one of embodiments 1-4, 6-9, 11-16, 18-19, 21, 26-27, 29, 31, or 33-34, wherein X ₂₀ is Aib. 1/GIP/Gcg receptor triple agonist.
36. The amino acid sequence of the peptide is YX ₂ QGTFTSDYS ₁₂ X ₁₃ LX ₁₅ KX ₁₇ X ₁₈ AX ₂₀ EFVX ₂₄ WLLX ₂₈ GGPSSGAPPPS (SEQ ID NO: 50),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
GLP-1/GIP/Gcg receptor according to any one of embodiments 1-3, 5-16, 18-19, 21, 26-27, or 29-32, wherein X ₂₈ is A or E. Triple agonist.
37. The amino acid sequence of the peptide is YX ₂ QGTFTSDYSX ₁₂ X ₁₃ LEKX ₁₇ AAX ₂₀ EFVQWLLEGGPSSGAPPPS (SEQ ID NO: 51),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₇ is Q or K,
GLP-1/GIP/ according to any one of embodiments 1-3, 5-9, 11-16, 18, 21, 26-27, 29, 31, or 36, wherein X ₂₀ is Aib. Gcg receptor triple agonist.
38. The amino acid sequence of the peptide is YX ₂ QGTFTSDYSX ₁₂ X ₁₃ LDKX ₁₇ AAX ₂₀ EFVNWLLAGGPSSGAPPPS (SEQ ID NO: 52),
X ₂ is Aib,
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₇ is Q or K,
GLP- according to any one of embodiments 1-3, 5-10, 12-13, 15-16, 18, 21, 26-27, 30, 32, or 36, wherein X ₂₀ is Aib. 1/GIP/Gcg receptor triple agonist.
39. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein the peptide is a C-terminal amide.
40. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 38, wherein the peptide is a C-terminal acid.
41. the substituent is ABC-,
A- is the following chemical formula 1
Chemical formula 1: HOOC-(CH ₂ ) _p -CO-*
(wherein p is an integer in the range of 16 to 20),
B- is the following chemical formula 2 or chemical formula 3
Chemical formula 2: *-NH-CH(COOH)-(CH ₂ ) ₂ -CO-*
Chemical formula 3: selected from *-NH-CH ₂ -(C ₆ H ₁₀ )-CO-NH-CH(COOH)-(CH ₂ ) ₂ -CO-*,
C- is the following chemical formula 4 or chemical formula 5
Chemical formula 4: *-[NH-((CH ₂ ) ₂ -O) ₂ -CH ₂ -CO] ₂ -*
Chemical formula 5: *-[NH-(CH ₂ ) ₄ -CH(NH ₂ )-CO] ₂ -*
as in any one of the preceding embodiments, wherein A-, B-, and C- are interconnected by an amide bond in the order indicated. GLP-1/GIP/Gcg receptor triple agonist.
42. The GLP-1/GIP/Gcg receptor triple agonist according to embodiment 41, wherein A- is Formula 1 and p is 16 or 18.
43. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 41-42, wherein B- is Formula 2.
44. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 41-42, wherein B- is Formula 3.
45. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 41-42 or 44, wherein B- is Formula 3a or Formula 3b.
46. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 41-45, wherein C- is Formula 4.
47. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 41-45, wherein C- is Formula 5.
48. The GLP according to any one of embodiments 41-43 or 46, wherein A- is Formula 1, p is 18, B- is Formula 2, and C- is Formula 4. -1/GIP/Gcg receptor triple agonist.
49. The GLP according to any one of embodiments 41-43 or 47, wherein A- is Formula 1, p is 18, B- is Formula 2, and C- is Formula 5. -1/GIP/Gcg receptor triple agonist.
50. Any one of the preceding embodiments, wherein the substituent ABC- is selected from the group consisting of Formula 6, Formula 7, Formula 8, Formula 9, Formula 10, Formula 11, Formula 12, and Formula 13. The GLP-1/GIP/Gcg receptor triple agonist described in .
51. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein the substituent ABC- is selected from the group consisting of Formula 12 and Formula 13.
52. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein the substituent is covalently bonded to the epsilon-amino group of K at position 16.
53. GLP-1/GIP/ according to any one of embodiments 1-15 or 18-51, wherein X ₁₇ is K and the substituent is covalently bonded to the epsilon-amino group of said K at position 17. Gcg receptor triple agonist.
54. The GLP according to any one of embodiments 1-26, 28-32, or 39-51, wherein X ₂₁ is K and the substituent is covalently bonded to the epsilon-amino group of said K at position 21. -1/GIP/Gcg receptor triple agonist.
55. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein the triple agonist is selected from the group consisting of Compound Nos. 1-57.
56. GLP-1/GIP/Gcg receptor according to any one of embodiments 1-52, or 55, wherein the triple agonist is selected from the group consisting of Compound Nos. 1, 5-6, and 12-57. Triple agonist.
57. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-51, 53, or 55, wherein the triple agonist is selected from the group consisting of Compound No. 2, and 9-10. medicine.
58. GLP-1/GIP/Gcg according to any one of embodiments 1-51, or 54-55, wherein the triple agonist is selected from the group consisting of compound numbers 3-4, 7-8, and 11. Triple receptor agonist.
59. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-52 or 55-56, wherein the triple agonist is Compound No. 26.
60. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-52 or 55-56, wherein the triple agonist is Compound No. 43.
61. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-52 or 55-56, wherein the triple agonist is Compound No. 44.
62. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, wherein the triple agonist is an agonist at hGLP-1R, hGIPR, and hGcgR.
63. A GLP-1/GIP/glucagon receptor triple agonist according to any one of the preceding embodiments, wherein the triple agonist is capable of activating hGLP-1R, hGIPR, and hGcgR.
64. as in any one of the preceding embodiments, wherein the triple agonist is capable of activating hGLP-1R, hGIPR, and hGcgR in an assay using whole cells expressing each of the three receptors. GLP-1/GIP/Gcg receptor triple agonist.
65. As described in any one of the preceding embodiments, wherein the triple agonist is capable of activating hGLP-1R, hGIPR, and hGcgR in vitro in a CRE luciferase assay such as Example 2 described herein. GLP-1/GIP/Gcg receptor triple agonist.
66. is an agonist at hGLP-1R with an EC ₅₀ of 50 pM or less, e.g. 10 pM or less, when measured in vitro without HSA, as determined in Example 2 herein. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the embodiments.
67. The preceding embodiment is an agonist at hGIPR with an EC ₅₀ of 50 pM or less, such as 20 pM or less, when measured in vitro without HSA, as determined in Example 2 herein. The GLP-1/GIP/Gcg receptor triple agonist according to any one of the above.
68. is an agonist at hGcgR with an EC ₅₀ of 500 pM or less, such as 250 pM or less, e.g. 50 pM or less, when measured in vitro without HSA, as determined in Example 2 herein. , a GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments.
69. hGLP-1R, hGIPR, and hGcgR all with an EC ₅₀ of 500 pM or less, e.g., 100 pM or less, when measured in vitro without HSA, as determined in Example 2 herein. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, which is an agonist.
70. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, having improved pharmacokinetic properties.
71. A GLP-1/GIP/Gcg receptor triple agonist according to any one of the preceding embodiments, having an increased half-life.
72. GLP-1/GIP/Gcg according to any one of the preceding embodiments, having an improved half-life when measured in minipigs, as determined in Example 3 herein. Triple receptor agonist.
73. according to any one of the preceding embodiments, having a half-life in minipigs of at least 30 hours, such as at least 40 hours, or at least 60 hours, as determined in Example 3 herein. GLP-1/GIP/Gcg receptor triple agonist.
74. GLP-1/GIP according to any one of the preceding embodiments, having in vivo efficacy in reducing food intake as determined in DIO mice, as in Example 4 herein. /Gcg receptor triple agonist.
75. GLP-1/GIP/Gcg reception according to any one of the preceding embodiments, with in vivo efficacy for inducing weight loss as determined in DIO mice, as in Example 4 herein. Triple agonist.
76. GLP-1/GIP/ according to any one of the preceding embodiments, having in vivo efficacy in improving glucose tolerance as determined in DIO mice, as in Example 4 herein. Gcg receptor triple agonist.
77. A pharmaceutical composition comprising a triple agonist according to any one of the preceding embodiments.
78. A pharmaceutical composition according to embodiment 77, and optionally at least one pharmaceutically acceptable excipient.
79. Pharmaceutical compositions according to embodiments 77-78, wherein the triple agonist is selected from the group consisting of Compound Nos. 1-57.
80. Pharmaceutical compositions according to embodiments 77-79, wherein the triple agonist is Compound No. 26.
81. Pharmaceutical compositions according to embodiments 77-79, wherein the triple agonist is Compound No. 43.
82. Pharmaceutical compositions according to embodiments 77-79, wherein the triple agonist is Compound No. 44.
83. A pharmaceutical composition according to any one of embodiments 77-82 for use as a medicament.
84. A GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for use as a medicament.
85. (i) Prevention and/or treatment of eating disorders such as obesity, e.g. by reducing food intake, increasing energy expenditure, reducing weight, suppressing appetite, inducing feelings of satiety; binge eating disorder, bulimia nervosa; treatment or prevention of obesity induced by the administration of antipsychotics or steroids, delayed gastric emptying, increased physical mobility, and/or comorbidities of obesity such as osteoarthritis and/or urinary incontinence. prevention and/or treatment of diseases;
(ii) weight maintenance after successful weight loss (either drug-induced or diet and exercise), i.e. prevention of weight gain after successful weight loss;
(iii) prevention and/or treatment of all forms of diabetes, including hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (young onset adult diabetes), gestational diabetes; and/or reduction of HbA1C,
(iv) slowing or preventing the progression of diabetic diseases, such as progression of type 2 diabetes, slowing the progression of impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, slowing or preventing insulin resistance, and/or or delaying the progression from type 2 diabetes that does not require insulin to type 2 diabetes that requires insulin;
(v) prevention of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver and/or A GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for use in the treatment of therapy.
86. Prevention and/or treatment of all forms of diabetes such as hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (adult onset diabetes of the young), gestational diabetes, and/or A GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-76 for use in reducing HbA1C.
87. Prevention and/or treatment of eating disorders such as obesity, binge eating disorder, bulimia nervosa, and/or by reducing food intake, increasing energy expenditure, reducing weight, suppressing appetite, and inducing satiety; Treatment or prevention of obesity induced by the administration of antipsychotics or steroids, slowing gastric emptying, increasing physical mobility, and/or preventing and/or co-morbidities of obesity such as osteoarthritis and/or urinary incontinence. or a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for use in therapy.
88. GLP according to any one of embodiments 1 to 76 for use in weight maintenance after successful weight loss (either drug-induced or diet and exercise) - i.e. preventing weight gain after successful weight loss. -1/GIP/Gcg receptor triple agonist.
89. Used for the prevention and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver. The GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1-76 for
90. GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for use in weight management, treatment and/or prevention of obesity and obesity-related disorders.
91. GLP-1/GIP/Gcg receptor according to any one of embodiments 1 to 76 for use in the treatment and/or prevention of all forms of diabetes, such as, for example, type 2 diabetes and diabetes-related disorders. Triple agonist.
92. (i) Prevention and/or treatment of eating disorders such as obesity, e.g. by reducing food intake, increasing energy expenditure, reducing weight, suppressing appetite, inducing feelings of satiety; binge eating disorder, bulimia nervosa; treatment or prevention of obesity induced by the administration of antipsychotics or steroids, delayed gastric emptying, increased physical mobility, and/or comorbidities of obesity such as osteoarthritis and/or urinary incontinence. prevention and/or treatment of diseases;
(ii) weight maintenance after successful weight loss (either drug-induced or diet and exercise), i.e. prevention of weight gain after successful weight loss;
(iii) prevention and/or treatment of all forms of diabetes, including hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (young onset adult diabetes), gestational diabetes; and/or reduction of HbA1C,
(iv) slowing or preventing the progression of diabetic diseases, such as progression of type 2 diabetes, slowing the progression of impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, slowing or preventing insulin resistance, and/or or delaying the progression from type 2 diabetes that does not require insulin to type 2 diabetes that requires insulin;
(v) prevention of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver and/or Use of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for the manufacture of a medicament for treatment.
93. Prevention and/or treatment of all forms of diabetes such as hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (adult onset diabetes of the young), gestational diabetes, and/or Use of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for the manufacture of a medicament for the reduction of HbA1C.
94. Prevention and/or treatment of eating disorders such as obesity, binge eating disorder, bulimia nervosa, and/or by reducing food intake, increasing energy expenditure, reducing weight, suppressing appetite, and inducing satiety; Treatment or prevention of obesity induced by the administration of antipsychotics or steroids, slowing gastric emptying, increasing physical mobility, and/or preventing and/or co-morbidities of obesity such as osteoarthritis and/or urinary incontinence. or for the manufacture of a medicament for treatment.
95. Any one of embodiments 1 to 76 for the manufacture of a medicament for weight maintenance after successful weight loss (either drug-induced or diet and exercise) - i.e. prevention of weight gain after successful weight loss. Use of the GLP-1/GIP/Gcg receptor triple agonist described in .
96. For the prevention and/or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver inflammation or fatty liver. Use of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for the manufacture of a medicament.
97. A GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76 for the manufacture of a medicament for weight management, treatment and/or prevention of obesity and obesity-related disorders. use.
98. GLP-1 according to any one of embodiments 1 to 76 for the manufacture of a medicament for the treatment and/or prevention of all forms of diabetes, such as type 2 diabetes and diabetes-related disorders. Use of GIP/Gcg receptor triple agonists.
99. e.g., reducing food intake, increasing energy expenditure, Prevention and/or treatment of eating disorders such as obesity by weight loss, suppression of appetite, and induction of satiety; binge eating disorder, bulimia nervosa, and/or induced by administration of antipsychotics or steroids Methods of treating or preventing obesity; slowing gastric emptying; increasing physical mobility; and/or preventing and/or treating comorbidities of obesity such as osteoarthritis and/or urinary incontinence.
100. (either drug-induced or diet and exercise) comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. Weight maintenance after successful weight loss - ie, how to prevent weight gain after successful weight loss.
101. Hyperglycemia, Type 2 Diabetes, Impaired Glucose Tolerance, 1, comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. Methods for the prevention and/or treatment of all forms of diabetes, such as type diabetes, non-insulin dependent diabetes, MODY (adult onset diabetes of the young), gestational diabetes, and/or for the reduction of HbA1C.
102. Delaying the progression of a diabetic disease, such as the progression of type 2 diabetes, comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. or prevention, delaying progression from impaired glucose tolerance (IGT) to type 2 diabetes requiring insulin, delaying or preventing insulin resistance, and/or from type 2 diabetes not requiring insulin to requiring insulin 2 How to slow progression to type diabetes.
103. hepatic steatosis, non-alcoholic steatosis, comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. A method for the prevention and/or treatment of liver disorders such as liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), hepatitis or fatty liver.
104. Weight management, treatment of obesity and obesity-related disorders and/or comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. Methods of prevention.
105. for example, type 2 diabetes and diabetes-related disorders, comprising administering a pharmaceutically active amount of a GLP-1/GIP/Gcg receptor triple agonist according to any one of embodiments 1 to 76. A method of treating and/or preventing forms of diabetes.
106. 77. A method for preparing a GLP-1/GIP/Gcg receptor triple agonist according to any of embodiments 1-76.

This experimental part begins with a list of abbreviations, followed by a section containing general methods for synthesizing and characterizing the triple agonists described herein. This is followed by some examples regarding the preparation of specific triple agonists and selected comparative compounds, and finally by some examples regarding the activity and properties of triple agonists.

The examples serve to illustrate the invention.

Abbreviation List The following abbreviations are in alphabetical order and are used below.
Ac: Acetyl Ado: 8-amino-3,6-dioxaoctanoic acid Aib: Alpha-aminoisobutyric acid (or α-aminoisobutyric acid)
amu: atomic mass unit API-ES: atmospheric pressure ionization-electrospray AUC: area under the curve BHK: baby hamster kidney Boc: tert-butyloxycarbonyl BW: body weight Cl-HOBt: 6-chloro-1-hydroxybenzotriazole CRE: cAMP response element DCM: dichloromethane DIC: diisopropylcarbodiimide DIPEA: N,N-diisopropylethylamine DMEM: Dulbecco's modified Eagle's medium DPBS: Dulbecco's phosphate buffered saline EDTA: ethylenediaminetetraacetic acid ELISA: enzyme-linked immunosorbent assay equiv: molar equivalents FBS: Fetal bovine serum Fmoc: 9-fluorenylmethyloxycarbonyl Gcg: Glucagon GcgR: Glucagon receptor GIP: Glucose-dependent insulinotropic polypeptide GIPR: Glucose-dependent insulinotropic polypeptide receptor GLP-1: Glucagon like peptide 1
GLP-1R: Glucagon-like peptide 1 receptor HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid HFIP: 1,1,1,3,3,3-hexafluoro-2-propanol or hexafluoro Isopropanol hGcgR: Human glucagon receptor hGIPR: Human glucose-dependent insulinotropic polypeptide receptor hGLP-1R: Human glucagon-like peptide 1 receptor HPLC: High performance liquid chromatography i. p. : Intraperitoneal IPGTT: Intraperitoneal glucose tolerance test i. v. : Intravenous LCMS: Liquid Chromatography Mass Spectrometry MeCN: Acetonitrile mM: mmol mmol: mmol min: min Mtt: 4-Methyltrityl MW: Molecular weight NMP: 1-methyl-pyrrolidin-2-one OtBu: tert-butyl ester Oxyma Pure (registered trademark): Cyano-hydroxyimino-acetic acid ethyl ester Pbf: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl PBS: Phosphate buffered saline PK: Pharmacokinetics pM: Picomole rpm: Number of rounds per minute Rt: Retention time SEM: Standard error SPPS: Solid phase peptide synthesis tBu: tert-butyl TFA: Trifluoroacetic acid TIS: Triisopropylsilane Trt: Triphenylmethyl or trityl Trx: Tranexamic acid TSTU: O- (N-succinimidyl)-1,1,3,3-tetramethyluronium tetrafluoroborate

General Preparation Methods Solid-phase peptide synthesis methods (including SPPS methods, deprotection of amino acids, cleavage of peptides from resins, and purification thereof), and methods of detection and characterization of the resulting peptides (LCMS method) is described below.

The resins used for the preparation of C-terminal peptide amides were H-Rink Amide-ChemMatrix resin (e.g., 0.5 mmol/g loading) or Rink Amide AM polystyrene resin (e.g., 0.6 mmol/g loading) or PAL Amide AM resin (eg, 0.6 mmol/g loading). The resin used for the preparation of C-terminal peptide acids was a Wang-polystyrene resin preloaded with the appropriate C-terminal Fmoc protected amino acids (eg, loaded at 0.6 mmol/g). Unless otherwise specified, the Fmoc-protected amino acid derivatives used were manufactured by, for example, AAPPTEC, Anaspec, Bachem, ChemImpex, Iris Biotech, Midwest Biotech, Gyros Protein Technologies, or Novabiochem. Fmoc-Ala-OH, Fmoc- Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)- OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc- Recommended standard products include Lys(Mtt)-OH and Fmoc-Aib-OH. The natural L form of the amino acid is used if nothing else is specified. The N-terminal amino acid was used with Boc protection at the alpha-amino group (e.g., Boc-Tyr(tBu)-OH for peptides with Tyr at the N-terminus, or Boc-Tyr(tBu)-OH for peptides with His at the N-terminus. His(Trt)-OH).

For modular albumin binding moiety attachment using SPPS, Fmoc-8-amino-3,6-dioxaoctanoic acid (Fmoc-Ado-OH), Fmoc-tranexamic acid (Fmoc-Trx-OH), Boc-Lys ( Fmoc)-OH, Fmoc-Glu-OtBu, octadecanedioic acid mono-tert-butyl ester, nonadecanedioic acid mono-tert-butyl ester, eicosanedioic acid mono-tert-butyl ester, or tetradecanedioic acid mono-tert-butyl ester Suitably protected building blocks were used, such as, but not limited to, esters. All operations described below were performed within the synthesis scale range of 0.1-0.5 mmol.

1. Synthesis of protected peptide backbone bound to resin:
Method: SPPS_A
SPPS was performed using Fmoc-based chemistry on a Protein Technologies SymphonyX solid-phase peptide synthesizer with some modifications to the protocol provided by the manufacturer. Mixing was accomplished by occasional nitrogen bubbling. The stepwise construction was carried out by the following steps: 1) pre-swelling the resin in DMF, 2) preparing the Fmoc in two treatments (10 min each) using 20% (v/v) piperidine in DMF. 3) Washing with DMF to remove piperidine; 4) Fmoc-amino acids (4-12 equivalents) and Oxyma Pure® (4-12 equivalents) each in 0.3-0.6 M DMF solutions; 12 eq.) followed by DIC (4-12 eq.) as a 0.6-1.2 M DMF solution and optionally DMF to adjust the final concentration of each component as needed. 4) washing with DMF to remove excess reagent; 5) final wash with DCM upon completion of construction. It was done using . Several amino acids were coupled with extended reaction times (eg, 4 hours) to ensure reaction completion, such as, but not limited to, a sterically hindered amino acid (eg, Aib) followed by an amino acid.

Method: SPPS_B
Protected peptidyl resins were synthesized according to the Fmoc strategy on an Applied Biosystems 431A solid phase peptide synthesizer using the general Fmoc protocol provided by the manufacturer. Mixing was achieved by vortexing and occasional nitrogen bubbling. The stepwise construction was carried out using the following steps: 1) Dissolving solid Fmoc-amino acid (10 eq.) in Cl-HOBt (10 eq.) as a 1M NMP solution, then adding DIC (10 eq.) as a 1M NMP solution. , then activating the Fmoc-amino acids by mixing simultaneously with steps 2-3, 2) a first treatment (3 min) using 20% (v/v) piperidine in NMP, then 3) Wash with NMP to remove piperidine; 4) Add activated Fmoc-amino acid solution to resin, then mix for 45-90 minutes. 4) Washing with NMP to remove excess reagents; 5) A final wash with DCM upon completion of the construction. The standard protected amino acid derivatives described above were supplied in pre-weighed cartridges (eg, from Midwest Biotech) and the non-standard derivatives were weighed manually. "double coupling" some amino acids, such as, but not limited to, a sterically hindered amino acid (e.g., Aib) followed by an amino acid, i.e., expelling the resin after the first coupling (e.g., 45 min). Completion of the reaction was ensured by adding further reagents (Fmoc-amino acids, DIC, Cl-HOBt) and allowing the mixture to react again (eg, 45 minutes).

2. Method for attaching substituents to resin-bound protected peptide skeleton: SC_A
N-epsilon-lysine protection The Mtt protecting group was removed by washing the resin with two treatments of 45 minutes each with 30% (v/v) HFIP in DCM, followed by washing with DCM and DMF. Acylation was performed with Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-tranexamic acid, Fmoc-Glu-OtBu, octadecanedioic acid mono-tert-butyl ester, and Performed on a Protein Technologies Symphony Ta.

Method: SC_B
N-epsilon-lysine protection The Mtt protecting group was removed by washing the resin with two treatments of 45 minutes each with 30% (v/v) HFIP in DCM, followed by washing with DCM and DMF. Acylation was performed with Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-tranexamic acid, Fmoc-Glu-OtBu, octadecanedioic acid mono-tert-butyl ester, and Performed on an Applied Biosystems 431A solid phase peptide synthesizer using the protocol described in Method SPPS_B using stepwise addition of building blocks such as, but not limited to, eicosanedioic acid mono-tert-butyl ester. Ta.

Method: SC_C
N-epsilon-lysine protection Mtt protecting group was removed by washing the resin with three treatments of 30 min each with 75:2.5:22.5 v/v/v HFIP/TIS/DCM followed by DCM and DMF. It was removed by washing with. Separately, 2-chlorotrityl chloride resin (e.g., loading 1.1 mmol/g) was transferred to a sintered separatory funnel and expanded in DCM for 30 min, then drained and placed in DCM (v/v). Shake with 10% DIPEA for 10 minutes. The resin was then drained and immediately treated with the first Fmoc protection building block of the substituents (1.0 eq.) and DIPEA (2 eq.) in DCM and shaken overnight at room temperature. Methanol was then added to the mixture to cap the unreacted sites of the resin. The resin was washed with DCM, DMF, and diethyl ether before drying under vacuum. Subsequently, the substituents were assembled stepwise as described in method SPPS_A. The protected substituent was cleaved from the resin using 20% HFIP (v/v) in DCM in three treatments of 10 min each, the cleavage mixture was concentrated in vacuo and dried under vacuo. This gave the protected substituent, which was used without further manipulation. The protected substituent was dissolved in dry THF at a concentration of 0.1 M before treatment with TSTU (1.5 eq.) and DIPEA (3 eq.). The mixture was stirred at room temperature for 2 hours, filtered and diluted 5 times with ethyl acetate. The organic extracts were washed with an equal volume of 0.1M aqueous HCl followed by an equal volume of water. The organic layer was then dried with MgSO ₄ and the volatile solvents were removed in vacuo. The activated substituent (1.5 eq.) was used directly as a 0.1 M solution in DMF containing DIPEA (3 eq.) and incubated with the peptidyl resin with free lysine ε-amino groups for 16 hours. The resin was then washed with DMF and DCM.

3. Cleavage and purification of resin-bound peptides:
After side chain synthesis was completed, the peptidyl resin was washed with DCM, dried, and then treated with TFA/water/TIS (95:2.5:2.5 (v/v/v)) for approximately 2-3 hours. , followed by precipitation with diethyl ether. The precipitate was washed with diethyl ether, dissolved in a suitable solvent (eg 2:1 water/MeCN) and left until all labile adducts decomposed. Purification is carried out, for example, by reverse phase preparative HPLC on a suitable column containing C8- or C18-silica gel (e.g. Waters 2545 binary gradient module, Waters 2489 UV/Visible detector, Waters Fraction Collector III, or e.g. Waters Deltaprep 4000). Separation of impurities and elution of product was achieved by increasing gradients of MeCN in water containing 0.1% TFA. Relevant fractions were checked for identity and purity by analytical LCMS. Fractions containing pure desired peptide were pooled and lyophilized to obtain the peptide TFA salt as a white solid.

4. Salt exchange from TFA to sodium salt:
3-20 mg of lyophilized purified peptide in a suitable aqueous buffer such as, but not limited to, 4:1 water/MeCN, 0.2 M sodium acetate, or 50 mM HEPES buffer pH 7.4. /mL. The pH of the solution was adjusted with aqueous NaOH as necessary to achieve complete solubility. The buffer containing the peptide was salt exchanged using a Sep-Pak C18 cartridge (0.5-5 g). The cartridge was equilibrated first with isopropanol, then with MeCN, then with water. The peptide solution was applied to the cartridge and the flow-through was reapplied to ensure complete retention of the peptide. The cartridge was washed with water and then with a buffer solution (eg, pH 7.5) including, but not limited to, NaHCO ₃ , NaOAc, or Na ₂ HPO ₄ . The cartridge was then washed with water and the peptides were eluted with 50-80% (v/v) MeCN in water. The eluate containing the peptide was lyophilized to yield the peptide sodium salt as a white solid, which was used as is.

Common detection and characterization methods LCMS methods:
Method: LCMS_A
LCMS_A was performed on a setup consisting of an Agilent 1260 Infinity series HPLC system and an Agilent Technologies 6120 Quadrupole MS. The eluents were defined as follows: A: 0.05% (v/v) TFA in water, B: 0.05% (v/v) in 9:1 (v/v) MeCN/water. T.F.A. The analysis was performed at a column temperature of 37° C. by injecting the appropriate volume of sample eluted with a gradient of A and B onto the column. Column: Phenomenex Kinetex C8, 2.6 μm, 100 Å, 4.6 x 75 mm. Gradient run time: Linear 20-100% B over 10 minutes at a flow rate of 1.0 mL/min. Detection: Diode array detector set at 214 nm. MS ionization mode: API-ES, positive polarity. MS scan mass range: 500-2000 amu. The most abundant isotope of each m/z is reported.

Method: LCMS_B
LCMS_B was performed on a setup consisting of a Waters Acquity H-class UPLC system and a Waters Xevo G2-XS QTof MS. The eluents were defined as follows: A: 0.1% (v/v) formic acid in water, B: 0.1% (v/v) formic acid in MeCN, C: 0.1% (v/v) formic acid in water. % (v/v) TFA. The analysis was performed at a column temperature of 40° C. by injecting the appropriate volume of sample eluted with a gradient of A and B onto the column. Column: Waters Acquity BEH, C-18, 1.7 μm, 2.1 x 50 mm. Gradient run time: Linear 5-95% B over 4.0 minutes at a flow rate of 0.4 mL/min in the presence of constant 5% C. MS ionization mode: ES, positive polarity. MS scan mass range: 50-4000 amu. The most abundant isotope of each m/z is reported.

[Example 1: Synthesis of compound]
The compounds are described below using the single letter amino acid code, with the exception of Aib. A substituent is included after the lysine (K) residue to which it is attached.

Compound number 1
Y-Aib-QGTFTSDYSIYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 1, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4821.3Da
LCMS_B: Retention time = 3.4 minutes, actual value [M+3H] ³⁺ 1607.1, [M+4H] ⁴⁺ 1205.6.

Compound number 2
Y-Aib-QGTFTSDYSIYLDK-K [2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 2, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K17
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4821.4Da
LCMS_B: Retention time = 3.0 minutes, actual values [M+2H] ²⁺ 2411.8, [M+3H] ³⁺ 1607.8, [M+4H] ⁴⁺ 1206.1.

Compound number 3
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 3, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K21
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4820.4Da
LCMS_B: Retention time = 3.0 minutes, actual value [M+3H] ³⁺ 1606.8, [M+4H] ⁴⁺ 1205.6.

Compound number 4
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 4, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K21
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4892.5Da
LCMS_A: Retention time = 6.1 min, actual value [M+3H] ³⁺ 1631.5, [M+4H] ⁴⁺ 1224.0.

Compound number 5
Y-Aib-QGTFTSDYSIYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 1, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 4849.4Da
LCMS_B: Retention time = 3.5 minutes, actual values [M+2H] ²⁺ 2424.7, [M+3H] ³⁺ 1616.5, [M+4H] ⁴⁺ 1212.6.

Compound number 6
Y-Aib-QGTFTSDYSYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 1, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 4988.6Da
LCMS_B: Retention time = 3.6 minutes, actual values [M+2H] ²⁺ 2495.1, [M+3H] ³⁺ 1663.4, [M+4H] ⁴⁺ 1247.8.

Compound number 7
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 4, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K21
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 4920.5Da
LCMS_B: Retention time = 3.2 minutes, actual value [M+2H] ²⁺ 2461.3, [M+3H] ³⁺ 1640.9.

Compound number 8
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynon decanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 4, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K21
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 5059.7Da
LCMS_B: Retention time = 3.3 minutes, actual value [M+3H] ³⁺ 1686.5, [M+4H] ⁴⁺ 1265.4.

Compound number 9
Y-Aib-QGTFTSDYSIYLDK-K [2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 2, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K17
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 4849.4Da
LCMS_B: Retention time = 3.2 minutes, actual value [M+H] ⁺ 4850.0 [M+2H] ²⁺ 2426.1, [M+3H] ³⁺ 1617.6.

Compound number 10
Y-Aib-QGTFTSDYSIYLDK-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 2, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K17
Synthesis method: SPPS_A; SC_C
Calculated molecular weight (average): 4988.6Da
LCMS_B: Retention time = 3.3 minutes, actual values [M+H] ⁺ 4988.0, [M+2H] ²⁺ 2495.4, [M+3H] ³⁺ 1663.6.

Compound 11
Y-Aib-QGTFTSDYSIYLEKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynon decanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 5, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K21
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 5073.7Da
LCMS_A: Retention time = 6.6 minutes, actual value [M+3H] ³⁺ 1692.0, [M+4H] ⁴⁺ 1269.4.

Compound number 12
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 6, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 5002.6Da
LCMS_A: Retention time = 6.9 minutes, actual value [M+3H] ³⁺ 1668.4, [M+4H] ⁴⁺ 1251.5.

Compound number 13
Y-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 6, C-terminal amide substituent: Chemical formula 11 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4968.7Da
LCMS_A: Retention time = 6.3 minutes, actual value [M+3H] ³⁺ 1657.0, [M+4H] ⁴⁺ 1243.1.

Compound number 14
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 7, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 5102.7Da
LCMS_A: Retention time = 7.0 minutes, actual value [M+3H] ³⁺ 1701.7, [M+4H] ⁴⁺ 1276.4.

Compound number 15
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 6, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4863.4Da
LCMS_A: Retention time = 6.9 minutes, actual value [M+3H] ³⁺ 1621.7, [M+4H] ⁴⁺ 1216.7.

Compound number 16
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 7, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4935.5Da
LCMS_A: Retention time = 7.0 minutes, actual value [M+3H] ³⁺ 1645.7, [M+4H] ⁴⁺ 1234.6.

Compound number 17
Y-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 7, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4901.5Da
LCMS_A: Retention time = 6.2 minutes, actual value [M+3H] ³⁺ 1634.5, [M+4H] ⁴⁺ 1226.7.

Compound number 18
Y-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 8, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4910.5Da
LCMS_A: Retention time = 6.1 min, actual value [M+3H] ³⁺ 1637.5, [M+4H] ⁴⁺ 1228.4.

Compound number 19
H-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 9, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4875.5Da
LCMS_A: Retention time = 7.0 minutes, actual value [M+3H] ³⁺ 1625.8, [M+4H] ⁴⁺ 1219.7.

Compound number 20
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAQEFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 10, C-terminal amide substituent: Chemical formula 10 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 5117.7Da
LCMS_A: Retention time = 6.8 minutes, actual value [M+3H] ³⁺ 1706.8, [M+4H] ⁴⁺ 1280.3.

Compound number 21
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 11, C-terminal amide substituent: Chemical formula 11 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 5023.7Da
LCMS_A: Retention time = 6.1 min, actual value [M+3H] ³⁺ 1675.3, [M+4H] ⁴⁺ 1256.7.

Compound number 22
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 11, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4884.5Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1628.8, [M+4H] ⁴⁺ 1222.0.

Compound number 23
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 12, C-terminal amide substituent: Chemical formula 11 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 5066.7Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1689.7, [M+4H] ⁴⁺ 1267.5.

Compound number 24
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 13, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4976.6Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1659.8. [M+4H] ⁴⁺ 1244.9.

Compound number 25
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 14, C-terminal amide substituent: Chemical formula 11 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 5073.7Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1692.0, [M+4H] ⁴⁺ 1269.1.

Compound number 26
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 14, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4934.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1645.5, [M+4H] ⁴⁺ 1234.5.

Compound number 27
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 15, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4928.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1643.6, [M+4H] ⁴⁺ 1232.9.

Compound number 28
H-Aib-HGTFTSDYSIYLD-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 16, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4870.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1624.4, [M+4H] ⁴⁺ 1218.5.

Compound number 29
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 17, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 5026.6Da
LCMS_A: Retention time = 5.8 minutes, actual value [M+3H] ³⁺ 1676.2, [M+4H] ⁴⁺ 1257.5.

Compound number 30
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 18, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4976.6Da
LCMS_B: Retention time = 2.7 minutes, actual value [M+3H] ³⁺ 1658.7, [M+4H] ⁴⁺ 1244.3.

Compound number 31
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 19, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4934.6Da
LCMS_B: Retention time = 2.7 minutes, actual value [M+3H] ³⁺ 1644.8, [M+4H] ⁴⁺ 1233.8.

Compound number 32
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 14, C-terminal amide substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 4906.5Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1636.4, [M+4H] ⁴⁺ 1227.3.

Compound number 33
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 20, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 4884.5Da
LCMS_A: Retention time = 5.7 minutes, actual value [M+3H] ³⁺ 1629.0, [M+4H] ⁴⁺ 1222.0.

Compound number 34
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 20, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 4918.5Da
LCMS_A: Retention time = 6.5 minutes, actual value [M+3H] ³⁺ 1640.3, [M+4H] ⁴⁺ 1230.4.

Compound number 35
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QKA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 21, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 4941.6Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1648.0, [M+4H] ⁴⁺ 1236.2.

Compound number 36
H-Aib-HGTFTSDYSKYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 22, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_B
Calculated molecular weight (average): 4899.5Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1633.8, [M+4H] ⁴⁺ 1225.7.

Compound number 37
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-RAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 23, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_B; SC_A
Calculated molecular weight (average): 4912.6Da
LCMS_A: Retention time = 5.8 minutes, actual value [M+3H] ³⁺ 1636.2, [M+4H] ⁴⁺ 1229.0.

Compound number 38
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 18, C-terminal amide substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4948.6Da
LCMS_A: Retention time = 5.5 minutes, actual value [M+3H] ³⁺ 1650.2, [M+4H] ⁴⁺ 1238.0.

Compound number 39
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 18, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 5010.6Da
LCMS_A: Retention time = 6.6 minutes, actual value [M+3H] ³⁺ 1670.9, [M+4H] ⁴⁺ 1253.7.

Compound number 40
H-Aib-HGTFTSDYSYYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 14, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4968.5Da
LCMS_A: Retention time = 6.5 minutes, actual value [M+3H] ³⁺ 1656.9, [M+4H] ⁴⁺ 1242.9.

Compound number 41
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 24, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4926.6Da
LCMS_A: Retention time = 5.8 minutes, actual value [M+3H] ³⁺ 1643.0, [M+4H] ⁴⁺ 1232.5.

Compound number 42
H-Aib-HGTFTSDYSYLLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 25, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4884.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1628.9, [M+4H] ⁴⁺ 1222.0.

Compound number 43
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 26, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4834.5Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1612.3, [M+4H] ⁴⁺ 1209.5.

Compound number 44
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 11, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4918.5Da
LCMS_A: Retention time = 6.7 minutes, actual value [M+3H] ³⁺ 1640.0, [M+4H] ⁴⁺ 1230.3.

Compound number 45
H-Aib-HGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 26, C-terminal amide substituent: Chemical formula 12 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4868.5Da
LCMS_A: Retention time = 6.7 minutes, actual value [M+3H] ³⁺ 1623.4, [M+4H] ⁴⁺ 1217.9.

Compound number 46
Y-Aib-QGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 27, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4851.5Da
LCMS_A: Retention time = 6.3 minutes, actual value [M+3H] ³⁺ 1617.8, [M+4H] ⁴⁺ 1213.6.

Compound number 47
Y-Aib-QGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 28, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4951.5Da
LCMS_A: Retention time = 6.2 minutes, actual value [M+3H] ³⁺ 1651.1, [M+4H] ⁴⁺ 1238.7.

Compound number 48
Y-Aib-QGTFTSDYSYLLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 29, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4901.5Da
LCMS_A: Retention time = 6.3 minutes, actual value [M+3H] ³⁺ 1634.5, [M+4H] ⁴⁺ 1226.3.

Compound number 49
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 30, C-terminal amide substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4878.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1626.9, [M+4H] ⁴⁺ 1220.3.

Compound number 50
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 30, C-terminal acid substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4879.5Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1627.0, [M+4H] ⁴⁺ 1220.8.

Compound number 51
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 30, C-terminal acid substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4851.4Da
LCMS_A: Retention time = 5.7 minutes, actual value [M+3H] ³⁺ 1617.8, [M+4H] ⁴⁺ 1213.7.

Compound number 52
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 31, C-terminal acid substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4850.4Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1617.5, [M+4H] ⁴⁺ 1213.4.

Compound number 53
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 31, C-terminal acid substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4878.5Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1626.8, [M+4H] ⁴⁺ 1220.3.

Compound number 54
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 32, C-terminal acid substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4878.5Da
LCMS_A: Retention time = 5.4 minutes, actual value [M+3H] ³⁺ 1626.8, [M+4H] ⁴⁺ 1220.4.

Compound number 55
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 32, C-terminal acid substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4878.5Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1636.2, [M+4H] ⁴⁺ 1227.4.

Compound number 56
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 33, C-terminal acid substituent: Chemical formula 7 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4859.5Da
LCMS_A: Retention time = 5.3 minutes, actual value [M+3H] ³⁺ 1620.4, [M+4H] ⁴⁺ 1215.7.

Compound number 57
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino )butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFVQWLLEGGPSSGAPPPS-OH
Peptide skeleton: SEQ ID NO: 33, C-terminal acid substituent: Chemical formula 13 bonded via the ε-amino group of K16
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4887.5Da
LCMS_A: Retention time = 5.6 minutes, actual value [M+3H] ³⁺ 1629.8, [M+4H] ⁴⁺ 1222.7.

Structures of standard and comparative compounds:
Native human GLP-1 (7-37), SEQ ID NO: 35
Native human GIP, SEQ ID NO: 36
Natural human glucagon, SEQ ID NO: 37
Native mouse GIP, SEQ ID NO: 34

Compound number 58
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-EFVNWLLAGGPSSGAPPPS-K[hexadecanoyl]-NH ₂
Peptide backbone: SEQ ID NO: 53, C-terminal amide substituent: hexadecanoyl bonded via the ε-amino group of K40 Synthesis method: See US9062124 Example 18, SEQ ID NO: 124 Calculated molecular weight (average): 4472. 1 Da
LCMS_A: Retention time = 7.1 min, actual value [M+3H] ³⁺ 1491.4, [M+4H] ⁴⁺ 1118.9.

Compound number 59
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-EFVNWLLAGGPSSGAPPPS-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl] _-NH2
Peptide skeleton: SEQ ID NO: 53, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K40
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4949.5Da
LCMS_A: Retention time = 6.1 min, actual value [M+3H] ³⁺ 1650.4, [M+4H] ⁴⁺ 1238.2.

Compound number 60
Y-Aib-QGTFTSDYSIYLDKQAA-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]--EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 38, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K20
Synthesis method: SPPS_A; SC_B
Calculated molecular weight (average): 4864.4Da
LCMS_A: Retention time = 6.1 min, actual value [M+3H] ³⁺ 1622.1, [M+4H] ⁴⁺ 1216.8.

Compound number 61
Y-Aib-QGTFTSDYS-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-YLDKQAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 39, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K12
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4836.4Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1612.7, [M+4H] ⁴⁺ 1210.0.

Compound number 62
Y-Aib-QGTFTSDYSI-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-LDKQAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 40, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K13
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4786.3Da
LCMS_A: Retention time = 6.0 minutes, actual value [M+3H] ³⁺ 1596.2, [M+4H] ⁴⁺ 1197.4.

Compound number 63
Y-Aib-QGTFTSDYSYSIY-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-DKQAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 41, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K14
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4836.4Da
LCMS_A: Retention time = 5.9 minutes, actual value [M+3H] ³⁺ 1613.0, [M+4H] ⁴⁺ 1209.9.

Compound number 64
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-EFV-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-WLLAGGPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 42, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K24
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4835.4Da
LCMS_A: Retention time = 6.2 minutes, actual value [M+3H] ³⁺ 1612.6, [M+4H] ⁴⁺ 1209.6.

Compound number 65
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-EFVNWLLA-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-GPSSGAPPPS-NH ₂
Peptide skeleton: SEQ ID NO: 43, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K29
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4892.5Da
LCMS_A: Retention time = 5.8 minutes, actual value [M+3H] ³⁺ 1631.5, [M+4H] ⁴⁺ 1223.9.

Compound number 66
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-EFVNWLLAGGPSSG-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino )butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-PPPS-NH ₂
Peptide skeleton: SEQ ID NO: 44, C-terminal amide substituent: Chemical formula 6 bonded via the ε-amino group of K35
Synthesis method: SPPS_A; SC_A
Calculated molecular weight (average): 4878.4Da
LCMS_A: Retention time = 6.2 minutes, actual value [M+3H] ³⁺ 1626.8, [M+4H] ⁴⁺ 1220.5.

[Example 2: In vitro functional efficacy (CRE luciferase, whole cells)]
The purpose of this example is to test the functional activity or efficacy of this compound in vitro at human and murine GLP-1, GIP, and Gcg receptors. In vitro functional efficacy is a measure of target receptor activation in whole cell assays. The potency of the compounds listed in Example 1 was determined as described below. Human GLP-1 (7-37) (SEQ ID NO: 35) (identical to mouse GLP-1 (7-37)), human GIP (SEQ ID NO: 36), and human glucagon (SEQ ID NO: 37) (identical to mouse glucagon) were included in the appropriate assay for comparison.

Rationale In vitro functional efficacy was determined by measuring target receptor responses in reporter gene assays in individual cell lines. This assay supports one of the following G protein-coupled receptors: human GLP-1 receptor, human GIP receptor, human Gcg receptor, mouse GLP-1 receptor, mouse GIP receptor, or mouse Gcg receptor. The experiments were carried out in stably transfected BHK cell lines expressing both cAMP response element (CRE) DNA and firefly luciferase (CRE luciferase) genes linked to a promoter. Activation of each receptor produces cAMP, which then results in the expression of luciferase protein. Once assay incubation is complete, luciferase substrate (luciferin) is added to cause enzymatic conversion of luciferin to oxyluciferin, resulting in bioluminescence. Luminescence was measured as the readout for this assay.

Cell Culture and Preparation The cell line used in these assays was BHK cells with BHKTS13 as the parental cell line. These cells were obtained from clones containing the CRE luciferase element and further established by transfecting the respective receptors to obtain related cell lines. The following cell lines were used:

Cells were cultured in cell culture medium at 37°C/5% _CO2 . These were aliquoted and stored in liquid nitrogen. These cells were cultured continuously and seeded the day before each assay.

Materials The following formulas were used in the assay: Pluronic F-68 10% (Gibco 2404), 10% fetal bovine serum (FBS, Invitrogen 16140-071), chicken egg white ovalbumin (Sigma A5503), phenol red-free DMEM (Gibco 21063-029), DMEM (Gibco 12430-054), 1M Hepes (Gibco 15630), Glutamax 100x (Gibco 35050), G418 (Invitrogen 10131-027), Hygromycin (Invitrogen en 10687-010), and steadylite plus (PerkinElmer 6016757 ).

Buffer GLP-1R and GcgR cell culture medium consisted of DMEM medium containing 10% FBS, 500 μg/mL G418, and 300 μg/mL hygromycin. GIPR cell culture medium consisted of DMEM medium containing 10% FBS, 400 μg/mL G418, and 300 μg/mL hygromycin. Assay buffer consisted of phenol red-free DMEM, 10 mM Hepes, 1x Glutamax, 1% ovalbumin, and 0.1% Pluronic F-68. Assay medium was mixed 1:1 with an equal volume of test compound in assay buffer to obtain the final assay concentration.

Procedure 1) Cells were plated at 5000 cells/well and incubated overnight in assay plates.
2) Cells were washed once with DPBS.
3) Test compound stocks and reference compound stocks at concentrations ranging from 100 to 300 μM were diluted 1:150 in assay buffer. Compounds were then diluted 1:10 in column 1 of a 96 deep-well dilution plate and then transferred to that row to create a 3.5-fold, 12-point dilution curve.
4) Assay buffer (50 μl aliquots) was added to each well of the assay plate.
5) Transfer a 50 μl aliquot of compound or blank from the dilution plate to the assay plate containing assay buffer.
6) Assay plates were incubated for 3 hours at 37°C in an incubator with 5% _CO2 .
7) Cells were washed once with DPBS.
8) Add 100 μl aliquots of DPBS to each well of the assay plate.
9) Added a 100 μl aliquot of steadylite plus reagent (photosensitive) to each well of the assay plate.
10) Cover each assay plate with aluminum foil to protect from light and shake at 250 RPM for 30 minutes at room temperature.
11) Each assay plate was read on a microtiter plate reader.

Calculations and Results Data from the microtiter plate reader was first regression analyzed in Excel to calculate the x-axis (log scale concentration) based on the stock concentration of each test compound and the dilution of the assay. This data was then transferred to GraphPad Prism software to generate graphs and perform statistical analysis. This software performs non-linear regression (log(agonist) versus response). The _EC50s calculated using this software and reported in pM are shown in Tables 1 and 2 below. A minimum of two replicates per sample were measured. Reported values are average values of replicates.

Compound No. 58 is known (Example 18 of US9062124, SEQ ID No. 124) and is a potent triple agonist with a hexadecanoyl moiety attached to the epsilon-amino group of Lys40. It was expected that replacing the substituent at Lys40 with substituent formula 6 (ie, Compound No. 59) would result in an equipotent compound with a longer half-life. However, the above results show that compound 59 has lower potency at all three receptors, especially GIPR and GcgR. Therefore, it is not possible to change the substituent from fatty monoacid to fatty acid diacid and obtain similar efficacy. The results in Table 1 indicate that placement of fatty acid diacid substituents on lysine is preferred when lysine is located at position 16, 17, or 21 of the peptide backbone, but the C18 diacid substituent chemical formula If 6 is in any of positions 12, 13, 14, 20, 24, 29, 35, or 40, a significant decrease in potency at one or more receptors is observed. It was done.

The results in Table 2 demonstrate that the compounds of the present invention exhibit potent functional activation of human GLP-1R, human GIPR, and human GcgR obtained by the peptide backbone in combination with substituents at positions 16, 17, or 21. Indicates that.

[Example 3: Pharmacokinetic study in minipigs]
The purpose of this example was to administer i.p. to minipigs. v. To determine the in vivo half-life of the triple agonists described herein after administration, ie, the prolongation of their time in the body and thus their time of action. This is done in pharmacokinetic (PK) studies to determine the terminal half-life of the triple agonist in question. Terminal half-life generally refers to the period of time it takes to halve a particular plasma concentration measured after the initial distribution phase.

Research Female Gottingen minipigs were obtained from Ellegaard Gottingen Minipigs (Dalmose, Denmark) and were used in this study, approximately 7-14 months old and weighing approximately 16-35 kg. Minipigs were individually housed and fed once a day on a restricted diet of SDS minipig diet (Special Diets Services, Essex, UK).

After 3 weeks of acclimatization, two permanent central venous catheters were implanted into the caudal tail of each minipig. Animals were allowed to recover for one week after surgery and then used for repeated pharmacokinetic studies with suitable washout periods between successive doses.

Minipigs were fasted approximately 18 hours before dosing and 0-4 hours after dosing, but had free access to water during the entire period.

The sodium salt of the triple agonist of Example 1 was prepared at a concentration of 20-40 nmol/mL in a buffer (pH 7.4) containing 0.025% (v/v) polysorbate 20, 10 mM sodium phosphate, 250 mM glycerol. It was dissolved in Intravenous injections of the triple agonist (usually in a volume corresponding to 1.5-2 nmol/kg, e.g. 0.1 mL/kg) are given through one catheter and blood samples are given at predetermined time points (preferably up to 14 days after administration). (through the other catheter). Blood samples (eg, 0.8 mL) were collected in 8 mM EDTA buffer and then centrifuged for 10 min at 4°C and 1942 g.

Sampling and Analysis Plasma was pipetted into Micronic tubes on dry ice and kept at −20° C. until analyzed for plasma concentrations of triple agonists using ELISA or similar antibody-based assays or LCMS. Individual plasma concentration-time profiles were analyzed using Phoenix WinNonLin ver. 6.4. (Pharsight Inc., Mountain View, CA, USA) and the resulting terminal half-life (harmonic mean) was determined.

result

The triple agonists tested herein have very long half-lives compared to natural hormones. The half-lives of hGLP-1 and hGIP measured in humans are reported to be approximately 2-4 minutes and 5-7 minutes, respectively (Meier et al., Diabetes, 2004, 53(3):654 -662). The half-life of glucagon measured in humans has been reported to vary between 5 and 30 minutes, depending on the route of administration (Pontiroli et al., Eur J Clin Pharmacol 1993, 45:555-558).

[Example 4: Pharmacodynamic research study in diet-induced obesity (DIO) mice]
The purpose of this example is to evaluate the in vivo effects of selected triple agonists on pharmacodynamic parameters in diet-induced obese (DIO) mice. Animals were treated once daily with a liquid formulation of the triple agonist tested by subcutaneous injection to assess the effects on body weight, paw intake, and glucose tolerance.

Animals and Diet C57BL/6J male mice were purchased from Jackson Laboratories at approximately 8 weeks of age. Mice were group housed and fed a high fat, high sugar diet from Research Diets (D12331). Mice were kept on this diet for 12 weeks before the start of the pharmacological study. Mice weighing more than 50 grams were considered diet-induced obese (DIO) and included in the pharmacological study. Mice were exposed to a controlled 12 h:12 h light/dark cycle at room temperature (22°C) and had free access to food and water. The study was approved by and conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee of the University of Cincinnati.

Dosing and Formulation All compounds in this study were formulated in the following buffer: 50mM phosphate, 70mM sodium chloride, 0.05% Tween-80, pH 7.4. Dosing solutions were formulated in glass vials and stored at 2-8°C. Dosing solutions were allowed to warm to room temperature before dosing and to 2-8°C after dosing.

DIO mice were distributed into groups (n=8/group) such that statistical variation in mean and standard deviation of fat mass and body weight was minimized between groups. Mice were grouped and treated as follows: vehicle or a GLP-1/GIP/Gcg receptor triple agonist as described herein (vehicle was 50 mM phosphate, 70 mM sodium chloride, 0.05 mg of phosphate, 70 mM sodium chloride; 05% (v/v) Tween-80, pH 7.4). Test compounds were dissolved in vehicle to a stock concentration of 100 μM and then diluted 50-200 times in vehicle to obtain the desired dosing solution concentration. Mice are dosed with the dosing solution in a volume of 2-5 μL per gram of body weight as needed to achieve the desired dose (e.g., 0.3 nmol/kg, 1.0 nmol/kg, or 3.0 nmol/kg). It was administered subcutaneously once a day in the morning.

Body weight and food intake Body weight (BW) and food intake were measured daily immediately before dosing. The percent body change for each mouse was calculated individually based on the initial body weight before the first injection.

IPGTT (intraperitoneal glucose tolerance test)
On the day of the glucose tolerance test, mice were fasted for 4 hours. Food was removed and mice were transferred to fresh cages. Mice had access to water but not food. Tail blood glucose levels were measured and mice were given an intraperitoneal (i.p.) glucose load of 2 g/kg (t=0) (glucose solution 200 mg/ml, dose volume 10 ml/kg). Tail blood glucose values were determined i. p. Measurements were taken at 0, 15, 30, 60, 90, and 120 minutes after glucose loading. Stratification of animals during IPGTT, e.g., administering 2 mice from group 1, followed by administering 2 mice from groups 2, 3, 4, then group 1, 2, 3 The next two mice were then dosed, and so on. This was to enable equal distribution of "time" across all groups.

result:
In one study, DIO mice received subcutaneous doses of vehicle, Compound No. 21, Compound No. 30, or Compound No. 33 daily for up to 60 days. The dose of each compound was titrated to maximum efficacy based on the titration schedule contained in Table 4. Mice that reached body weight normalization, defined as a weight of 22.5 g or less, were voluntarily removed from the study. The results are shown in Table 5 and Figures 1-2. All triple agonist compounds tested reduced body weight and food intake to a greater extent than vehicle. At time points prior to day 42 (see Figure 1), treatment with Compound No. 21 resulted in a similar reduction in food intake compared to Compound No. 30 and Compound No. 33, but body weight The reduction was inferior. This shows that the similar GLP-1R of compound no. 21 compared to compound no. 30 and compound no. and the in vitro efficacy of GIPR may be explained by the decreased efficacy of GcgR in vitro. In vitro efficacy at mouse GLP-1R, mouse GIPR, and mouse GcgR is shown in Table 6.

Additional mouse studies using Compound Nos. 26, 43, and 44 demonstrated superior reductions in food intake and body weight compared to vehicle in DIO mice after daily subcutaneous administration for 13 days. The results are shown in Table 7. Compound No. 44 showed dose-dependent effects on reducing food intake and body weight and improving glucose tolerance compared to vehicle at 1.0 nmol/kg and 3.0 nmol/kg doses. This indicates that in triple agonists, weight loss efficacy can be increased without deleteriously affecting glucose tolerance.

While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will occur to those skilled in the art. It is therefore to be understood that the appended claims are intended to cover all modifications and changes falling within the true spirit of the invention.

Claims (15)

A GLP-1/GIP/Gcg receptor triple agonist comprising a peptide and a substituent, wherein the amino acid sequence of the peptide is
_{X 1 X 2} X _{3 GTFTSDYS 12} X _{13 LX 15} KX _{17 X 18} AX 20
X ₂ is Aib,
X ₃ is H or Q,
X ₁₂ is K, I, or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q, R, or K,
X ₁₈ is A, Y, or K,
X ₂₀ is Aib, Q, E, R, or H;
X ₂₁ is E or K,
X ₂₄ is N or Q,
X ₂₈ is A or E,
the peptide is a C-terminal acid or amide,
the substituent is covalently bonded to the epsilon-amino group of K at a position selected from the group consisting of positions 16, 17, and 21;
A GLP-1/GIP/Gcg receptor triple agonist, or a pharmaceutically acceptable salt thereof. The GLP-1/GIP/Gcg receptor triple agonist according to claim 1, wherein the substituent is covalently bonded to the epsilon-amino group of K at position 16. The GLP-1/GIP/Gcg receptor triple agonist according to claim 1 or 2, wherein X ₁ X ₂ X ₃ is selected from the group consisting of HAibH and YAibQ. _{SX 12 _} , the GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 3. _5. The method _of claims 1-4, wherein KX ₁₇ GLP-1/GIP/Gcg receptor triple agonist according to any one of the above. The amino acid sequence of the peptide is
_{HX 2 HGTFTSDYSX 12 _ _ _ _ _ _}
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 5, wherein X ₂₈ is A or E. The amino acid sequence of the peptide is
_{YX 2 QGTFTSDYS 12 X 13 LX 15 KX 17}
X ₁₂ is I or Y,
X ₁₃ is Y or L,
X ₁₅ is D or E,
X ₁₇ is Q or K,
X ₁₈ is A or Y,
X ₂₀ is Aib,
X ₂₄ is N or Q,
GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 5, wherein X ₂₈ is A or E. The substituent is ABC-, in which A- is represented by the following chemical formula 1
Chemical formula 1: HOOC-(CH ₂ ) _p -CO-*,
p is an integer in the range of 16 to 20,
B- is the following chemical formula 2 or chemical formula 3
Chemical formula 2: *-NH-CH(COOH)-(CH ₂ ) ₂ -CO-*
Chemical formula 3: selected from *-NH-CH ₂ -(C ₆ H ₁₀ )-CO-NH-CH(COOH)-(CH ₂ ) ₂ -CO-*,
C- is the following chemical formula 4 or chemical formula 5
Chemical formula 4: *-[NH-((CH ₂ ) ₂ -O) ₂ -CH ₂ -CO] ₂ -*
Chemical formula 5: selected from *-[NH-(CH ₂ ) ₄ -CH(NH ₂ )-CO] ₂ -*,
GLP-1/GIP according to any one of claims 1 to 7, wherein * indicates the point of attachment and A, B and C are interconnected by amide bonds in the order indicated. /Gcg receptor triple agonist. The substituent ABC- is
and
GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 8, selected from the group consisting of. The triple agonist is
Y-Aib-QGTFTSDYSIYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 1):
Y-Aib-QGTFTSDYSIYLDK-K [2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 2):
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 3):
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 4):
Y-Aib-QGTFTSDYSIYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 5):
Y-Aib-QGTFTSDYSYLD-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 6):
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl ]Amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 7):
Y-Aib-QGTFTSDYSIYLDKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynon decanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂ (compound number 8):
Y-Aib-QGTFTSDYSIYLDK-K [2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 9):
Y-Aib-QGTFTSDYSIYLDK-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl Amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-AA-Aib-EFVNWLLAGGPSSGAPPPS- _NH2 (Compound No. 10):
Y-Aib-QGTFTSDYSIYLEKQAA-Aib-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynon decanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-FVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 11):
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 12):
Y-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 13):
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 14):
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVNWLLAGGPSSGAPPPS-NH ₂ (Compound No. 15):
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 16):
Y-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 17):
Y-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 18):
H-Aib-QGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 19):
Y-Aib-QGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-[[4-[(19-carboxynonadecanoyl amino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAQEFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 20):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 21):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 22):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 23):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 24):
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-[[4-[(19- carboxynonadecanoylamino)methyl]cyclohexanecarbonyl]amino]butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 25):
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 26):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 27):
H-Aib-HGTFTSDYSIYLD-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 28):
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 29):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) Butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 30):
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 31):
H-Aib-HGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 32):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 33):
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-KAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 34):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QKA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (compound number 35):
H-Aib-HGTFTSDYSKYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 36):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-RAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 37):
H-Aib-HGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 38):
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 39):
H-Aib-HGTFTSDYSYYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 40):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) Butanoyl]amino]hexanoyl]amino]hexanoyl]-KYA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 41):
H-Aib-HGTFTSDYSYLLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 42):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 43):
H-Aib-HGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 44):
H-Aib-HGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino ]Ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 45):
Y-Aib-QGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 46):
Y-Aib-QGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 47):
Y-Aib-QGTFTSDYSYLLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAA-Aib-EFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 48):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-NH ₂ (Compound No. 49):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-OH (compound number 50):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFVQWLLEGGPSSGAPPPS-OH (compound number 51):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-OH (compound number 52):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVQWLLEGGPSSGAPPPS-OH (compound number 53):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVQWLLEGGPSSGAPPPS-OH (compound number 54):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVQWLLEGGPSSGAPPPS-OH (compound number 55):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFVQWLLEGGPSSGAPPPS-OH (compound number 56):
H-Aib-HGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino ) butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFVQWLLEGGPSSGAPPPS-OH (compound number 57):
GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 9, selected from the group consisting of. A pharmaceutical composition comprising a GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 10 and optionally at least one pharmaceutically acceptable excipient. thing. GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 10 for use as a medicament. GLP-1/GIP/Gcg receptor triple agonist according to any one of claims 1 to 10 for use in weight management, treatment and/or prevention of obesity and obesity-related disorders. GLP-1/GIP/Gcg receptor according to any one of claims 1 to 10 for use in the treatment and/or prevention of all forms of diabetes, such as type 2 diabetes, and diabetes-related disorders. Triple agonist. Claim 1 for use in the treatment and/or prevention of liver disorders, such as hepatic steatosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver inflammation, or fatty liver. The GLP-1/GIP/Gcg receptor triple agonist according to any one of items 1 to 10.